Modulation of acetylcholine level by donepezil hydrochloride attenuates the severity of experimental autoimmune encephalomyelitis (EAE), an animal model for multiple sclerosis

Multiple sclerosis (MS), is a chronic autoimmune disease characterized by neuroinflammation and demyelination, with no known cure. Previous studies demonstrated that crotoxin, a neurotoxin derived from rattlesnake venom, effectively alleviated hyperalgesia and clinical symptoms in experimental autoimmune encephalomyelitis (EAE) when combined with silica SBA-15 (CTX-SBA-15), an inert nanostructured particle. This combination inhibited disease progression and reduced neuroinflammation. Building on these findings, we conducted a comprehensive gene expression analysis to identify differentially expressed genes (DEGs) and pathways affected by MOG_35–55-induced EAE and modulated by CTX, which could serve as potential targets for disease control. Gene expression in the central nervous system was analyzed using the GeneChip platform. Data were processed using the Transcriptome Analysis Console and the Gene Ontology database. The analysis identified 1055 DEGs, with 238 genes specifically associated with the disease in the EAE vs. CFA comparison. In the CTX-SBA-15 post-treatment group, 695 DEGs were identified, with 204 genes upregulated and 491 downregulated compared to the EAE group. Overrepresentation analysis highlighted acetylcholine receptors pathways as significant targets. For validation, donepezil hydrochloride, an acetylcholinesterase inhibitor, was administered (2 mg/kg/day) initiating 12 days after immunization, with body weight monitored every other day. Donepezil improved clinical symptoms, reduced weight loss, decreased cellular infiltrates, and prevented spinal cord demyelination. In conclusion, our findings highlights that the acetylcholine pathway represents a promising target to be modulated for MS management.